“…Promoter CpG island methylation of genes reported to be methylated in CRC (2,5,22,23): mutL homolog1, colon cancer, nonpolyposis type 2 (Escherichia coli; MLH1), cyclin-dependent kinase inhibitor 2A (CDKN2A; p16INK4 and p14ARF), O-6-methylguanine-DNA methyltransferase (MGMT), Ras association (RalGDS/AF-6) domain family member 1 (RASSF1A), adenomatous polyposis coli (APC), helicase-like transcription factor (HLTF), GATA-binding protein 4 (GATA4), GATA-binding protein 5 (GATA5), checkpoint with forkhead and ring finger domains (CHFR), ADAM metallopeptidase domain 23 (ADAM23), Rab32, member RAS oncogene family (RAB32), junctophilin (JPH3), forkhead box L2 (FOXL2), BCL2-adenovirus E1B 19kDA interacting protein 3 (BNIP3), neutralized homolog (Drosophila; NEURL), calcium channel, voltage dependent, a2-delta subunit 1 (CACNA2), thrombospondin 1 (THBS1), tissue factor pathway inhibitor 2 (TFPI2), and the CIMP genes calcium channel, voltage-dependent, T type, a-1G subunit (CACNA1G), insulin-like growth factor-II (somatomedin A, IGF-II), neurogenin 1 (NEUROG1), runt-related transcription factor 3 (RUNX3), and suppressor of cytokine signaling 1 (SOCS1) were determined using sodium bisulfite modification of genomic DNA (EZ DNA Methylation Kit, ZYMO research Co.). To facilitate methylation-specific PCR (MSP) analysis on DNA retrieved from formalinfixed, paraffin-embedded tissue, nested MSP was performed as described elsewhere (24,25).…”